An optimized method for establishing high purity murine CD8(+) T cell cultures

作者：Zanker Damien; Xiao Kun; Oveissi Sara; Guillaume Philippe; Luescher Immanuel F; Chen Weisan^*

来源：Journal of Immunological Methods, 2013, 387(1-2): 173-180.

DOI：10.1016/j.jim.2012.10.012

• CD8(+) T cells
• T cell culture
• B220
• Apoptosis

Establishing CD8(+) T cell cultures has been empirical and the published methods have been largely individual laboratory based. In this study, we optimized culturing conditions and show that IL-2 concentration is the most critical factor for the success of establishing CD8(+) T cell cultures. High IL-2 concentration encouraged T cells to non-specifically proliferate, express a B cell marker, B220, and undergo apoptosis. These cells also lose typical irregular T cell morphology and are incapable of sustaining long-term cultures. Using tetramer and intracellular cytokine assessments, we further demonstrated that many antigen-specific T cells have been rendered nonfunctional when expanded under high IL-2 concentration. When IL-2 is used in the correct range, B220-mediated cell depletion greatly enhanced the success rate of such T cell cultures.

• 出版日期2013-1